One thousand milliliter of sample was introduced into a given beaker containing 100.0 μg of hexahydrothymol
as internal standard. 10% NaOH was employed to adjust the pH (PH≥1 1).Basic contents of the sample were then extracted 3 times by 50 ml of dichloromethane.The pH of the remaining liquor w as adjusted to a pH o f 2 u sing 6 mol/L H 2SO4, an d acidic contents were extracted as described above. The two extracted liquors were brought together and dried for about 24 h by anhydrous sulfate sodium, and finally evaporated to 1.0ml in K-D (Kudema-Danish evaporator). 1.0 μ1 of the remaining liquor was withdrawn for GC-MS analysis.
Finnigan Voyager was used as analytical tool for gas chromatography-mass spectrometry (GC-MS) analysis, with DB-5 silica capillary column (30 m x 0.25 mm, 0.25 μm). The column temperature was programmed from 50℃ (for 2 min) to 300℃ (for 10min) with a heating-up velocity of 10}C /min, carrier gas of He was 1.0 ml/min, and a split ratio of 20:1. Source temperature was held at 200`C, ionization energy at 70 eV (electron volt). Interpretation of the
resulting chromatograms and mass spectra was performed using NIST (National Institute of Standards and Technology) library.